INHIBITORS OF p38

ABSTRACT

The present invention relates to inhibitors of p38, a mammalian protein kinase involved cell proliferation, cell death and response to extracellular stimuli. The invention also relates to inhibitors of ZAP70. The invention also relates to methods for producing these inhibitors. The invention also provides pharmaceutical compositions comprising the inhibitors of the invention and methods of utilizing those compositions in the treatment and prevention of various disorders.

TECHNICAL FIELD OF INVENTION

The present invention relates to inhibitors of p38, a mammalian proteinkinase involved in cell proliferation, cell death and response toextracellular stimuli. The invention also relates to methods forproducing these inhibitors. The invention also provides pharmaceuticalcompositions comprising the inhibitors of the invention and methods ofutilizing those compositions in the treatment and prevention of variousdisorders.

BACKGROUND OF THE INVENTION

Protein kinases are involved in various cellular responses toextracellular signals. Recently, a family of mitogen-activated proteinkinases (MAPK) has been discovered. Members of this family are Ser/Thrkinases that activate their substrates by phosphorylation [B. Stein etal., Ann. Rep. Med. Chem., 31, pp. 289-98 (1996)]. MAPKs are themselvesactivated by a variety of signals including growth factors, cytokines,UV radiation, and stress-inducing agents.

One particularly interesting MAPK is p38. p38, also known as cytokinesuppressive anti-inflammatory drug binding protein (CSBP) and RK, wasisolated from murine pre-B cells that were transfected with thelipopolysaccharide (LPS) receptor, CD14, and induced with LPS. p38 hassince been isolated and sequenced, as has the cDNA encoding it in humansand mouse. Activation of p38 has been observed in cells stimulated bystress, such as treatment of lipopolysaccharides (LPS), UV, anisomycin,or osmotic shock, and by cytokines, such as IL-1 and TNF.

Inhibition of p38 kinase leads to a blockade on the production of bothIL-1 and TNF. IL-1 and TNF stimulate the production of otherproinflammatory cytokines such as IL-6 and IL-8 and have been implicatedin acute and chronic inflammatory diseases and in post-menopausalosteoporosis [R. B. Kimble et al., Endocrinol., 136, pp. 3054-61(1995)].

Based upon this finding, it is believed that p38, along with otherMAPKs, have a role in mediating cellular response to inflammatorystimuli, such as leukocyte accumulation, macrophage/monocyte activation,tissue resorption, fever, acute phase responses and neutrophilia. Inaddition, MAPKs, such as p38, have been implicated in cancer,thrombin-induced platelet aggregation, immunodeficiency disorders,autoimmune diseases, cell death, allergies, osteoporosis andneurodegenerative disorders. Inhibitors of p38 have also been implicatedin the area of pain management through inhibition of prostaglandinendoperoxide synthase-2 induction. Other diseases associated with Il-1,IL-6, IL-8 or TNF overproduction are set forth in WO 96/21654.

Others have already begun trying to develop drugs that specificallyinhibit MAPKs. For example, PCT publication WO 95/31451 describespyrazole compounds that inhibit MAPKs, and, in particular, p38. However,the efficacy of these inhibitors in vivo is still being investigated.

Other p38 inhibitors have been produced, including those described in WO98/27098, WO 99/00357, WO 99/10291, WO 99/58502, WO 99/64400, WO00/17175 and WO 00/17204.

Accordingly, there is still a great need to develop other potentinhibitors of p38, including p38-specific inhibitors, that are useful intreating various conditions associated with p38 activation.

Another protein kinase that is involved in cellular responses toextracellular signals is ZAP70. When the T cell receptor (TCR) in Tcells is triggered by binding an antigen, it in turn activates ZAP70.ZAP70 acts to couple the TCR to a number of essential signallingpathways that are required for T cell differentiation and proliferation.

Given ZAP70's role in T cell signalling, ZAP70 may have a role in T cellmediated diseases. Such diseases include, without limitation,transplantation, autoimune disease, e.g., RA, systemic lupuserythematosus (SLE), psoriasis, Sjogren's Syndrome, thyroiditis,pulmonary fibrosis, bronchiolitis obliterans, hemolytic anemia andWegener's granulomatosis, cancer, including leukemia and lymphoma,multiple sclerosis, graft versus host disease, and Kawasaki syndrome.

Accordingly, there is a great need to develop inhibitors of ZAP70 thatare useful in treating various conditions associated with ZAP70activation.

SUMMARY OF THE INVENTION

The present invention addresses this problem by providing compounds thatdemonstrate inhibition of p38 and/or ZAP70.

These compounds have the general formula:

wherein each of Q₁ and Q₂ are independently selected from a phenyl or5-6 membered aromatic heterocyclic ring system, or a 8-10 memberedbicyclic ring system comprising aromatic carbocyclic rings, aromaticheterocyclic rings or a combination of an aromatic carbocyclic ring andan aromatic heterocyclic ring.

A heterocyclic ring system or a heterocyclic ring contains 1 to 4heteroatoms, which are independently selected from N, O, S, SO and SO₂.

The rings that make up Q₁ are substituted with 1 to 4 substituents, eachof which is independently selected from halo; C₁-C₃ alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; O—(C₁-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; NR′₂; OCF₃; CF₃; NO₂;CO₂R′; CONR′; SR′; S(O₂)N(R′)₂; SCF₃; CN; N(R′)C(O)R⁴; N(R′)C(O)OR⁴;N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴; N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂;or N═C—N(R′)₂.

The rings that make up Q₂ are optionally substituted with up to 4substituents, each of which is independently selected from halogen;C₁-C₃ straight or branched alkyl optionally substituted with R′, NR′₂,OR′, CO₂R′, S(O₂)N(R′)₂, N═C—N(R′)₂, R³, O—P(O₃)H₂, or CONR′₂;O—(C₁-C₃)-alkyl; O—(C₁-C₃)-alkyl optionally substituted with NR′₂, OR′,CO₂R′, S(O₂)N(R′)₂, N═CR′—N(R′)₂, R³, OP(O₃)H₂, or CONR′₂; NR′₂; OCF₃;CF₃; NO₂; CO₂R′; CONR′₂; R³; OR³; NR³ ₂; SR³; C(O)R³; C(O)N(R′)R³;C(O)OR³; SR′; S(O₂)N(R′)₂; SCF₃; N═CR′—N(R′)₂; OR⁴; O—CO₂R⁴;N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴;N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; K; or CN.

Each R′ is independently selected from hydrogen; (C₁-C₃)-alkyl;(C₂-C₃)-alkenyl or alkynyl; phenyl or phenyl substituted with 1 to 3substituents independently selected from halo, methoxy, cyano, nitro,amino, hydroxy, methyl or ethyl; or a 5-6 membered heterocyclic ringsystem optionally substituted with 1 to 3 substituents independentlyselected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl orethyl.

Each R is independently selected from hydrogen, —R², —N(R²)₂, —OR², SR²,—C(O)—N(R²)₂, —S(O₂)—N(R²)₂, —C(O)—OR² or —C(O)R² wherein two adjacent Rare optionally bound to one another and, together with each Y to whichthey are respectively bound, form a 4-8 membered carbocyclic orheterocyclic ring.

Each R² is independently selected from hydrogen; or (C₁-C₃)-alkyl or(C₁-C₃)-alkenyl, each optionally substituted with —N(R′)₂, —OR′, SR′,—O—C(O)—N(R′)₂, —C(O)—N(R′)₂, —S(O₂)—N(R′)₂, —C(O)—OR′, —NSO₂R⁴,—NSO₂R³, —C(O)N(R′)(R³), —NC(O)R⁴, —N(R′)(R³), —N(R′)(R⁴), —C(O)R³,—C(O)N(R′)(R⁴), —N(R⁴)₂, —C(O)N═C(NH)₂ or R³.

Each R³ is independently selected from 5-8 membered aromatic ornon-aromatic carbocyclic or heterocyclic ring systems each optionallysubstituted with R′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or —K; or an 8-10membered bicyclic ring system comprising aromatic carbocyclic rings,aromatic heterocyclic rings or a combination of an aromatic carbocyclicring and an aromatic heterocyclic ring each optionally substituted withR′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or —K.

Each R⁴ is independently selected from R′; (C₁-C₇)-straight or branchedalkyl optionally substituted with R′, N(R′)₂, OR′, CO₂R′, CON(R′)₂,SO₂N(R′)₂ or SO₂N(R⁵)₂; or a 5-6 membered carbocyclic or heterocyclicring system optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂,SO₂N(R′)₂ or SO₂N(R⁵)₂.

Each R⁵ is independently selected from hydrogen, (C₁-C₃)-alkyl, or(C₁-C₃)-alkenyl; each optionally substituted with —N(R′)₂, —OR′, SR′,—C(O)—N(R′)₂, —S(O₂)—N(R′)₂, —C(O)—OR′, —N—S(O₂)(R′), —NSO₂R⁶,—C(O)N(R′)(R⁶), —NC(O)R′, —N(R′)(R⁶), —C(O)R⁶, —C(O)N═C(NH)₂ or R⁶.

Each R⁶ is independently selected from 5-8 membered aromatic ornon-aromatic carbocyclic or heterocyclic ring systems each optionallysubstituted with R′, —C(O)R′ or —C(O)OR′; or an 8-10 membered bicyclicring system comprising aromatic carbocyclic rings, aromatic heterocyclicrings or a combination of an aromatic carbocyclic ring and an aromaticheterocyclic ring each optionally substituted with R′, —C(O)R′ orC(O)OR′.

R⁷ is selected from H, halogen, or a (C₁-C₃) straight chain or branchedalkyl.

Each Y is independently selected from N or C. If either Y is N, then Ror U attached to Y is a lone pair of electrons.

Z is CH, N, C(OCH₃), C(CH₃), C(NH₂), C(OH) or C(F).

Each U is independently selected from R or J.

Each J is independently selected from a (C₁-C₄) straight chain orbranched alkyl derivative substituted with T.

Each T is independently selected from either O(V) or N(H)(V).

Each V is independently selected from C(O)N═C(R)(N(R)₂), wherein the twogeminal R on the nitrogen are optionally bound to one another to form a4-8 membered carbocyclic or heterocyclic ring.

When the two R components form a ring, it will obvious to those skilledin the art that a terminal hydrogen from each unfused R component willbe lost. For example, if a ring structure is formed by binding those twoR components together, one being —CH₃ and the other being —CH₂—CH₃, oneterminal hydrogen on each R component (indicated in bold) will be lost.Therefore, the resulting portion of the ring structure will have theformula —CH₂—CH₂—CH₂—.

Each K is independently selected from a (C₁-C₄) straight chain orbranched alkyl derivative substituted with D, or —OP(O)(OH)₂.

Each D is independently selected from either enantiomer of

Each M is independently selected from either O or NH.

Each G is independently selected from NH₂, OH, or H.

Each R₈ is independently selected from H, OH, C(O)OH, (C₁-C₇)-straightor branched alkyl optionally substituted with N(R′)₂, OR′, CO₂R′,CON(R′)₂, or SO₂N(R⁵)₂; or a 5-6 membered carbocyclic, heterocyclic orheteroaryl ring system optionally substituted with N(R′)₂, OR′, CO₂R′,CON(R′)₂, or SO₂N(R⁵)₂. When G forms a ring with R₈, it will be obviousto those skilled in the art that a terminal hydrogen from the unfused Gand R₈ component will be lost. For example, if a ring structure isformed by binding the G and R₈ components together, one being —NH₂ andthe other being —CH₂—CH₂—CH₂—CH₃, one terminal hydrogen on each Rcomponent (indicated in bold) will be lost. Therefore, the resultingportion of the ring structure will have the formula—NH—CH₂—CH₂—CH₂—CH₂—.

In another embodiment, the invention provides pharmaceuticalcompositions comprising the p38 and/or ZAP70 inhibitors of thisinvention. These compositions may be utilized in methods for treating orpreventing a variety of p38-mediated disorders, such as cancer,inflammatory diseases, autoimmune diseases, destructive bone disorders,proliferative disorders, infectious diseases, viral diseases andneurodegenerative diseases or ZAP70-mediated disorders, includingtransplantation, autoimune disease, cancer, multiple sclerosis, graftversus host disease, and Kawasaki syndrome. These compositions are alsouseful in methods for preventing cell death and hyperplasia andtherefore may be used to treat or prevent reperfusion/ischemia instroke, heart attacks, and organ hypoxia. The compositions are alsouseful in methods for preventing thrombin-induced platelet aggregation.Each of these above-described methods is also part of the presentinvention.

DETAILED DESCRIPTION OF THE INVENTION

These compounds have the general formula:

wherein each of Q₁ and Q₂ are independently selected from a phenyl or5-6 membered aromatic heterocyclic ring system, or a 8-10 memberedbicyclic ring system comprising aromatic carbocyclic rings, aromaticheterocyclic rings or a combination of an aromatic carbocyclic ring andan aromatic heterocyclic ring.

The rings that make up Q₁ are substituted with 1 to 4 substituents, eachof which is independently selected from halo; C₁-C₃ alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; O—(C₁-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; NR′₂; OCF₃; CF₃; NO₂;CO₂R′; CONR′; SR′; S(O₂)N(R′)₂; SCF₃; CN; N(R′)C(O)R⁴; N(R′)C(O)OR⁴;N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴; N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂;or N═C—N(R′)₂.

The rings that make up Q₂ are optionally substituted with up to 4substituents, each of which is independently selected from halogen;C₁-C₃ straight or branched alkyl optionally substituted with R′, NR′₂,OR′, CO₂R′, S(O₂)N(R′)₂, N═C—N(R′)₂, R³, O—P(O₃)H₂, or CONR′₂;O—(C₁-C₃)-alkyl; O—(C₁-C₃)-alkyl optionally substituted with NR′₂, OR′,CO₂R′, S(O₂)N(R′)₂, N═CR′—N(R′)₂, R³, OP(O₃)H₂, or CONR′₂; NR′₂; OCF₃;CF₃; NO₂; CO₂R′; CONR′₂; R³; OR³; NR³ ₂; SR³; C(O)R³; C(O)N(R′)R³;C(O)OR³; SR′; S(O₂)N(R′)₂; SCF₃; N═CR′—N(R′)₂; OR⁴; O—CO₂R⁴;N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴;N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; K; or CN.

Each R′ is independently selected from hydrogen; (C₁-C₃)-alkyl;(C₂-C₃)-alkenyl or alkynyl; phenyl or phenyl substituted with 1 to 3substituents independently selected from halo, methoxy, cyano, nitro,amino, hydroxy, methyl or ethyl; or a 5-6 membered heterocyclic ringsystem optionally substituted with 1 to 3 substituents independentlyselected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl orethyl.

Each R is independently selected from hydrogen, —R², —N(R²)₂, —OR², SR²,—C(O)—N(R²)₂, —S(O₂)—N(R²)₂, —C(O)—OR² or —C(O)R² wherein two adjacent Rare optionally bound to one another and, together with each Y to whichthey are respectively bound, form a 4-8 membered carbocyclic orheterocyclic ring.

Each R² is independently selected from hydrogen; or (C₁-C₃)-alkyl or(C₁-C₃)-alkenyl, each optionally substituted with —N(R′)₂, —OR′, SR′,—O—C(O)—N(R′)₂, —C(O)—N(R′)₂, —S(O₂)—N(R′)₂, —C(O)—OR′, —NSO₂R⁴,—NSO₂R³, —C(O)N(R′)(R³), —NC(O)R⁴, —N(R′)(R³), —N(R′)(R⁴), —C(O)R³,—C(O)N(R′)(R⁴), —N(R⁴)₂, —C(O)N═C(NH)₂ or R³.

Each R³ is independently selected from 5-8 membered aromatic ornon-aromatic carbocyclic or heterocyclic ring systems each optionallysubstituted with R′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or —K; or an 8-10membered bicyclic ring system comprising aromatic carbocyclic rings,aromatic heterocyclic rings or a combination of an aromatic carbocyclicring and an aromatic heterocyclic ring each optionally substituted withR′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or —K.

Each R⁴ is independently selected from R′; (C₁-C₇)-straight or branchedalkyl optionally substituted with R′, N(R′)₂, OR′, CO₂R′, CON(R′)₂,SO₂N(R′)₂ or SO₂N(R⁵)₂; or a 5-6 membered carbocyclic or heterocyclicring system optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂,SO₂N(R′)₂ or SO₂N(R⁵)₂.

Each R⁵ is independently selected from hydrogen, (C₁-C₃)-alkyl, or(C₁-C₃)-alkenyl; each optionally substituted with —N(R′)₂, —OR′, SR′,—C(O)—N(R′)₂, —S(O₂)—N(R′)₂, —C(O)—OR′, —N—S(O₂)(R′), —NSO₂R⁶,—C(O)N(R′)(R⁶), —NC(O)R′, —N(R′)(R⁶), —C(O)R⁶, —C(O)N═C(NH)₂ or R⁶.

Each R⁶ is independently selected from 5-8 membered aromatic ornon-aromatic carbocyclic or heterocyclic ring systems each optionallysubstituted with R′, —C(O)R′ or —C(O)OR′; or an 8-10 membered bicyclicring system comprising aromatic carbocyclic rings, aromatic heterocyclicrings or a combination of an aromatic carbocyclic ring and an aromaticheterocyclic ring each optionally substituted with R′, —C(O)R′ orC(O)OR′.

R⁷ is selected from H, halogen, or a (C₁-C₃) straight chain or branchedalkyl.

Each Y is independently selected from N or C. If either Y is N, then Ror U attached to Y is a lone pair of electrons.

Z is CH, N, C(OCH₃), C(CH₃), C(NH₂), C(OH) or C(F).

Each U is independently selected from R or J.

Each J is independently selected from a (C₁-C₄) straight chain orbranched alkyl derivative substituted with T.

Each T is independently selected from either O(V) or N(H)(V).

Each V is independently selected from C(O)N═C(R)(N(R)₂), wherein the twogeminal R on the nitrogen are optionally bound to one another to form a4-8 membered carbocyclic or heterocyclic ring.

When the two R components form a ring, it will obvious to those skilledin the art that a terminal hydrogen from each unfused R component willbe lost. For example, if a ring structure is formed by binding those twoR components together, one being —CH₃ and the other being —CH₂—CH₃, oneterminal hydrogen on each R component (indicated in bold) will be lost.Therefore, the resulting portion of the ring structure will have theformula —CH₂—CH₂—CH₂—.

Each K is independently selected from a (C₁-C₄) straight chain orbranched alkyl derivative substituted with D, or —OP(O)(OH)₂.

Each D is independently selected from either enantiomer of

Each M is independently selected from either O or NH.

Each G is independently selected from NH₂, OH, or H.

Each R₈ is independently selected from H, OH, C(O)OH, (C₁-C₇)-straightor branched alkyl optionally substituted with N(R′)₂, OR′, CO₂R′,CON(R′)₂, or SO₂N(R⁵)₂; or a 5-6 membered carbocyclic, heterocyclic orheteroaryl ring system optionally substituted with N(R′)₂, OR′, CO₂R′,CON(R′)₂, or SO₂N(R⁵)₂. When G forms a ring with R₈, it will be obviousto those skilled in the art that a terminal hydrogen from the unfused Gand R₈ component will be lost. For example, if a ring structure isformed by binding the G and R₈ components together, one being —NH₂ andthe other being —CH₂—CH₂—CH₂—CH₃, one terminal hydrogen on each Rcomponent (indicated in bold) will be lost. Therefore, the resultingportion of the ring structure will have the formula—NH—CH₂—CH₂—CH₂—CH₂—.

A heterocyclic ring system or a heterocyclic ring contains 1 to 4heteroatoms, which are independently selected from N, O, and S. Asubstitutable nitrogen on an aromatic or non-aromatic heterocyclic ringmay be optionally substituted. N or S may also exist in oxidized formsuch as NO, SO and SO₂.

One having ordinary skill in the art will recognize that the maximumnumber of heteroatoms in a stable, chemically feasible heterocyclicring, whether it is aromatic or non-aromatic, is determined by the sizeof the ring, degree of unsaturation, and valence of the heteroatoms. Ingeneral, a heterocyclic ring may have one to four heteroatoms so long asthe heterocyclic ring is chemically feasible and stable.

The term “chemically stable arrangement” or “chemically feasible andstable” as used herein, refers to a compound structure that renders thecompound sufficiently stable to allow manufacture and administration toa mammal by methods known in the art. Typically, such compounds arestable at a temperature of 40° C. or less, in the absence of moisture orother chemically reactive conditions, for at least a week.

According to a preferred embodiment, Q₁ is selected from phenyl orpyridyl containing 1 to 3 substituents, wherein at least one of saidsubstituents is in the ortho position and said substituents areindependently selected from chloro, fluoro, bromo, —CH₃, —OCH₃, —OH,—CF₃, —OCF₃, —O(CH₂)₂CH₃, NH₂, 3,4-methylenedioxy, —N(CH₃)₂,—NH—S(O)₂-phenyl, —NH—C(O)O—CH₂-4-pyridine, —NH—C(O)CH₂-morpholine,—NH—C(O)CH₂—N(CH₃)₂, —NH—C(O)CH₂-piperazine, —NH—C(O)CH₂-pyrrolidine,—NH—C(O)C(O)-morpholine, —NH—C(O)C(O)-piperazine,—NH—C(O)C(O)-pyrrolidine, —O—C(O)CH₂—N(CH₃)₂, or —O—(CH₂)₂—N(CH₃)₂.

Even more preferred are phenyl or pyridyl containing at least 2 of theabove-indicated substituents both being in the ortho position.

Some specific examples of preferred Q₁ are:

Most preferably, Q₁ is selected from 2-fluoro-6-trifluoromethylphenyl,2,6-difluorophenyl, 2,6-dichlorophenyl, 2-chloro-4-hydroxyphenyl,2-chloro-4-aminophenyl, 2,6-dichloro-4-aminophenyl,2,6-dichloro-3-aminophenyl, 2,6-dimethyl-4-hydroxyphenyl,2-methoxy-3,5-dichloro-4-pyridyl, 2-chloro-4,5methylenedioxy phenyl, or2-chloro-4-(N-2-morpholino-acetamido)phenyl.

According to a preferred embodiment, Q₂ is phenyl, pyridyl or naphthylcontaining 0 to 3 substituents, wherein each substituent isindependently selected from chloro, fluoro, bromo, methyl, ethyl,isopropyl, —OCH₃, —OH, —NH₂, —CF₃, —OCF₃, —SCH₃, —OCH₃, —C(O)OH,—C(O)OCH₃, —CH₂NH₂, —N(CH₃)₂, —CH₂-pyrrolidine and —CH₂OH.Some specific examples of preferred Q₂ are:

unsubstituted 2-pyridyl or unsubstituted phenyl.

Most preferred are compounds wherein Q₂ is selected from phenyl,2-isopropylphenyl, 3,4-dimethylphenyl, 2-ethylphenyl, 3-fluorophenyl,2-methylphenyl, 3-chloro-4-fluorophenyl, 3-chlorophenyl,2-carbomethoxylphenyl, 2-carboxyphenyl, 2-methyl-4-chlorophenyl,2-bromophenyl, 2-pyridyl, 2-methylenehydroxyphenyl, 4-fluorophenyl,2-methyl-4-fluorophenyl, 2-chloro-4-fluorphenyl, 2,4-difluorophenyl,2-hydroxy-4-fluorphenyl, 2-methylenehydroxy-4-fluorophenyl, 1-naphthyl,3-chloro-2-methylenehydroxy, 3-chloro-2-methyl, or 4-fluoro-2-methyl.

According to another preferred embodiment, R⁷ is a halogen. In a morepreferred embodiment, R⁷ is Cl.

According to another preferred embodiment, each Y is C.

According an even more preferred embodiment, each Y is C and the R and Uattached to each Y component is hydrogen.

Some specific examples of preferred J are:

According to another preferred embodiment, K is a 0-4 atom chainterminating in an ester.

According to another preferred embodiment, M is O.

Some specific examples of preferred K are:

More preferably, K is selected from:

Some preferred embodiments are provided in Tables 1 to 3 below: TABLE 1Cmpd Nmbr Structure 101

102

103

104

105

106

107

108

109

110

TABLE 2 Cmpd Nmbr Structure 111

112

113

114

115

116

117

TABLE 3 Cmpd Nmbr Structure 118

119

120

121

122

123

124

125

Particularly preferred embodiments include:

wherein Ar is

Particularly preferred embodiments also include:

wherein Ar is

Other particularly preferred embodiments include:

wherein Ar is

Other particularly preferred embodiments include:

wherein

Other particularly preferred embodiments include:

wherein

Other particularly preferred embodiments include:

wherein X is N(CH₃)₂,

Other particularly preferred embodiments include:

wherein Y=Me or H; and X=(CH₂)₃, CH₂C(CH₃)₂CH₂, CH₂N(Me)C(O)CH₂.

Some most preferred embodiments include:

According to another embodiment, the present invention provides methodsof producing the above-identified compounds of the formulae (Ia), (Ib),(Ic) or (Id). Representative synthesis schemes are depicted below. Inall schemes, the L1 and L2 groups on the initial materials are meant torepresent leaving groups ortho to the nitrogen atom in a heterocyclicring. For example, compound A may be 2,6-dichloro-3 nitro pyridine.

One having skill in the art will recognize Scheme 1 may be used tosynthesize compounds having the general formula of (Ia), (Ib), (Ic) and(Id).

According to another embodiment of the invention, the activity of thep38 inhibitors of this invention may be assayed in vitro, in vivo or ina cell line. In vitro assays include assays that determine inhibition ofeither the kinase activity or ATPase activity of activated p38.Alternate in vitro assays quantitate the ability of the inhibitor tobind to p38 and may be measured either by radiolabelling the inhibitorprior to binding, isolating the inhibitor/p38 complex and determiningthe amount of radiolabel bound, or by running a competition experimentwhere new inhibitors are incubated with p38 bound to known radioligands.

Cell culture assays of the inhibitory effect of the compounds of thisinvention may determine the amounts of TNF, IL-1, IL-6 or IL-8 producedin whole blood or cell fractions thereof in cells treated with inhibitoras compared to cells treated with negative controls. Level of thesecytokines may be determined through the use of commercially availableELISAs.

An in vivo assay useful for determining the inhibitory activity of thep38 inhibitors of this invention are the suppression of hind paw edemain rats with Mycobacterium butyricum-induced adjuvant arthritis. This isdescribed in J. C. Boehm et al., J. Med. Chem., 39, pp. 3929-37 (1996),the disclosure of which is herein incorporated by reference. The p38inhibitors of this invention may also be assayed in animal models ofarthritis, bone resorption, endotoxin shock and immune function, asdescribed in A. M. Badger et al., J. Pharmacol. ExperimentalTherapeutics, 279, pp. 1453-61 (1996), the disclosure of which is hereinincorporated by reference.

The p38 inhibitors or pharmaceutical salts thereof may be formulatedinto pharmaceutical compositions for administration to animals orhumans. These pharmaceutical compositions, which comprise an amount ofp38 inhibitor effective to treat or prevent a p38-mediated condition anda pharmaceutically acceptable carrier, are another embodiment of thepresent invention.

The term “p38-mediated condition”, as used herein means any disease orother deleterious condition in which p38 is known to play a role. Thisincludes conditions known to be caused by IL-1, TNF, IL-6 or IL-8overproduction. Such conditions include, without limitation,inflammatory diseases, autoimmune diseases, destructive bone disorders,proliferative disorders, infectious diseases, neurodegenerativediseases, allergies, reperfusion/ischemia in stroke, heart attacks,angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiachypertrophy, thrombin-induced platelet aggregation, and conditionsassociated with prostaglandin endoperoxidase synthase-2.

Inflammatory diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, acute pancreatitis,chronic pancreatitis, asthma, allergies, and adult respiratory distresssyndrome.

Autoimmune diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, glomerulonephritis,rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmunehemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopicdermatitis, chronic active hepatitis, myasthenia gravis, multiplesclerosis, inflammatory bowel disease, ulcerative colitis, Crohn'sdisease, psoriasis, or graft vs. host disease.

Destructive bone disorders which may be treated or prevented by thecompounds of this invention include, but are not limited to,osteoporosis, osteoarthritis and multiple myeloma-related bone disorder.

Proliferative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, acutemyelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma,Kaposi's sarcoma, and multiple myeloma.

Angiogenic disorders which may be treated or prevented by the compoundsof this invention include solid tumors, ocular neovasculization,infantile haemangiomas.

Infectious diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, sepsis, septic shock,and Shigellosis.

Viral diseases which may be treated or prevented by the compounds ofthis invention include, but are not limited to, acute hepatitisinfection (including hepatitis A, hepatitis B and hepatitis C), HIVinfection and CMV retinitis.

Neurodegenerative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, Alzheimer'sdisease, Parkinson's disease, cerebral ischemias or neurodegenerativedisease caused by traumatic injury.

“p38-mediated conditions” also include ischemia/reperfusion in stroke,heart attacks, myocardial ischemia, organ hypoxia, vascular hyperplasia,cardiac hypertrophy, and thrombin-induced platelet aggregation.

In addition, p38 inhibitors of the instant invention are also capable ofinhibiting the expression of inducible pro-inflammatory proteins such asprostaglandin endoperoxide synthase-2 (PGHS-2), also referred to ascyclooxygenase-2 (COX-2). Therefore, other “p38-mediated conditions”which may be treated by the compounds of this invention include edema,analgesia, fever and pain, such as neuromuscular pain, headache, cancerpain, dental pain and arthritis pain.

The diseases that may be treated or prevented by the p38 inhibitors ofthis invention may also be conveniently grouped by the cytokine (IL-1,TNF, IL-6, IL-8) that is believed to be responsible for the disease.

Thus, an IL-1-mediated disease or condition includes rheumatoidarthritis, osteoarthritis, stroke, endotoxemia and/or toxic shocksyndrome, inflammatory reaction induced by endotoxin, inflammatory boweldisease, tuberculosis, atherosclerosis, muscle degeneration, cachexia,psoriatic arthritis, Reiter's syndrome, gout, traumatic arthritis,rubella arthritis, acute synovitis, diabetes, pancreatic β-cell diseaseand Alzheimer's disease.

TNF-mediated disease or condition includes, rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, gouty arthritis and otherarthritic conditions, sepsis, septic shock, endotoxic shock, gramnegative sepsis, toxic shock syndrome, adult respiratory distresssyndrome, cerebral malaria, chronic pulmonary inflammatory disease,silicosis, pulmonary sarcoidosis, bone resorption diseases, reperfusioninjury, graft vs. host reaction, allograft rejections, fever andmyalgias due to infection, cachexia secondary to infection, AIDS, ARC ormalignancy, keloid formation, scar tissue formation, Crohn's disease,ulcerative colitis or pyresis. TNF-mediated diseases also include viralinfections, such as HIV, CMV, influenza and herpes; and veterinary viralinfections, such as lentivirus infections, including, but not limited toequine infectious anemia virus, caprine arthritis virus, visna virus ormaedi virus; or retrovirus infections, including feline immunodeficiencyvirus, bovine immunodeficiency virus, or canine immunodeficiency virus.

IL-8 mediated disease or condition includes diseases characterized bymassive neutrophil infiltration, such as psoriasis, inflammatory boweldisease, asthma, cardiac and renal reperfusion injury, adult respiratorydistress syndrome, thrombosis and glomerulonephritis.

In addition, the compounds of this invention may be used topically totreat or prevent conditions caused or exacerbated by IL-1 or TNF. Suchconditions include inflamed joints, eczema, psoriasis, inflammatory skinconditions such as sunburn, inflammatory eye conditions such asconjunctivitis, pyresis, pain and other conditions associated withinflammation.

According to another embodiment, the compounds of this invention may beused to treat ZAP70-mediated conditions including, without limitation,organ or tissue rejection associated with transplantation, autoimunedisease, e.g., rheumatoid arthritis, systemic lupus erythematosus (SLE),psoriasis, Sjogren's Syndrome, thyroiditis, pulmonary fibrosis,bronchiolitis obliterans, hemolytic anemia and Wegener's granulomatosis,cancer, including leukemia and lymphoma, multiple sclerosis, graftversus host disease, and Kawasaki syndrome.

The ZAP70 inhibitors or pharmaceutical salts thereof may be formulatedinto pharmaceutical compositions for administration to animals orhumans. These pharmaceutical compositions, which comprise an amount ofZAP70 inhibitor effective to treat or prevent a ZAP70-mediated conditionand a pharmaceutically acceptable carrier, are another embodiment of thepresent invention.

In addition to the compounds of this invention, pharmaceuticallyacceptable salts of the compounds of this invention may also be employedin compositions to treat or prevent the above-identified disorders.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of suitable acid salts includeacetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptanoate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, salicylate, succinate, sulfate, tartrate,thiocyanate, tosylate and undecanoate. Other acids, such as oxalic,while not in themselves pharmaceutically acceptable, may be employed inthe preparation of salts useful as intermediates in obtaining thecompounds of the invention and their pharmaceutically acceptable acidaddition salts. Salts derived from appropriate bases include alkalimetal (e.g., sodium and potassium), alkaline earth metal (e.g.,magnesium), ammonium and N—(C1-4 alkyl)4+ salts. This invention alsoenvisions the quaternization of any basic nitrogen-containing groups ofthe compounds disclosed herein. Water or oil-soluble or dispersibleproducts may be obtained by such quaternization.

Pharmaceutically acceptable carriers that may be used in thesepharmaceutical compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

The amount of p38 or ZAP70 inhibitor that may be combined with thecarrier materials to produce a single dosage form will vary dependingupon the host treated and the particular mode of administration.Preferably, the compositions should be formulated so that a dosage ofbetween 0.01-100 mg/kg body weight/day of the inhibitor can beadministered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of inhibitor will also depend upon the particular compound in thecomposition.

According to another embodiment, the invention provides methods fortreating or preventing a p38-mediated condition comprising the step ofadministering to a patient one of the above-described pharmaceuticalcompositions. The term “patient”, as used herein, means an animal,preferably a human.

Preferably, that method is used to treat or prevent a condition selectedfrom inflammatory diseases, autoimmune diseases, destructive bonedisorders, proliferative disorders, infectious diseases, degenerativediseases, allergies, reperfusion/ischemia in stroke, heart attacks,angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiachypertrophy, and thrombin-induced platelet aggregation.

According to another embodiment, the inhibitors of this invention areused to treat or prevent an IL-1, IL-6, IL-8 or TNF-mediated disease orcondition. Such conditions are described above.

Depending upon the particular p38-mediated condition to be treated orprevented, additional drugs, which are normally administered to treat orprevent that condition, may be administered together with the inhibitorsof this invention. For example, chemotherapeutic agents or otheranti-proliferative agents may be combined with the p38 inhibitors ofthis invention to treat proliferative diseases.

Those additional agents may be administered separately, as part of amultiple dosage regimen, from the p38 inhibitor-containing composition.Alternatively, those agents may be part of a single dosage form, mixedtogether with the p38 inhibitor in a single composition.

According to another embodiment, the invention provides methods fortreating or preventing a ZAP70-mediated condition comprising the step ofadministering to a patient one of the above-described pharmaceuticalcompositions.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXAMPLE 1 Synthesis of p38 Inhibitor Compound 7

To a solution of LDA (60 mmol, 40 mLs) at −78° C., was added dropwise asolution of 2,6-dibromopyridine (40 mmol, 9.48 gms) in THF (30 mLs,dried). The mixture was stirred at −78° C. for 20 minutes. Ethyl formate(400 mmol, 32.3 mLs) was added and stirring was continued at −78° C. for2 hours. Saturated ammonium chloride (200 mLs) was added and the mixturewas warmed to room temperature. The reaction mixture was diluted withethyl acetate and the organic layer was washed with aqueous acid andbase. The organic layer was dried and evaporated in vacuo. The resultingmaterial was purified by flash chromatography on silica gel followed byeluting with 10% ethyl acetate in n-hexane to afford 1 (32 mmol, 8.41gms) as a white solid.

A solution of 1 (776 mmol, 205.6 gms) and triethyl orthoformate (200 mL)dissolved in ethanol (750 mL) was refluxed overnight. The reactionmixture was cooled, and evaporated in vacuo. The remaining red oil wasdissolved in hexane and filtered over a plug of silica gel. The plug waseluted with 50% CH₂Cl₂/hexane. The filtrate was evaporated to afford 2as an oil.

To a suspension of 60% NaH (130 mmol, 5.20 g) and 2 (61.2 mmol, 20.76 g)in THF (100 mL) at reflux was added dropwise a solution of2,6-difluoroaniline (61.3 mmol, 20 g) in THF (100 mL). After the anilinehad been added, Pd(PPh₃)₄ (100 mg) was added. The mixture was refluxedfor one hour and cooled. Hydrochloric acid (1N, 100 mL) was added andstirring was continued for one hour. The reaction mixture was extractedwith CH₂Cl₂. The organic extract was dried and evaporated in vacuo. Theresulting material was dissolved in a minimal amount of CH₂Cl₂ andhexane was added. The solution was cooled precipitating 3 as a yellowsolid.

p-fluorophenylboronic acid (57.5 mmol, 8.05 g), and 3 (46.9 mmol, 14.70g) were dissolved in a dimethoxyethane (300 mL). Cesium fluoride (68.6mmol, 10.42 g) and tetrakis(triphenylphosphine)palladium (0) (100 mgs)were added to the solution and the suspension was allowed to refluxovernight. The reaction mixture was poured into water and extracted withCH₂Cl₂. The organic extract was washed with 1N NaOH, dried with MgSO₄,and filtered over a plug of silica gel. The plug was eluted with CH₂Cl₂and the filtrate was evaporated in vacuo. The resulting yellow solid wastriturated with 50% CH₂Cl₂/hexane to afford 4 (9.50 g, 62%) as a yellowsolid.

A solution of 4 (70.1 mmol, 23.01 g) in toluene (250 mL) was combinedwith a 20% solution of phosgene in toluene (151 mmol, 80 mL) and heatedto reflux for two hours. The reaction was cooled and poured intoammonium hydroxide. The mixture was stirred for five minutes andextracted with methylene chloride. The organic extract was dried andfiltered over a plug of silica gel. The plug was eluted with methylenechloride to remove residual starting material. It was then eluted with50% ethyl acetate/methylene chloride to obtain 5. The filtrate wasevaporated in vacuo to afford 5 (21.38 g, 86%) as a white solid.

Sodium borohydride (36.5 mmol, 1.38 g) was added to a solution of 5(60.0 mmol, 21.38 g) in THF (100 mL) and the solution was stirred forone hour at 0° C. and then two hours at room temperature. The reactionwas poured into 1N HCl and extracted with methylene chloride. Theorganic extract was dried and filtered over a plug of silica gel. Theplug was eluted with 5% ethyl acetate/methylene chloride to removeresidual starting material. It was then eluted with ethyl acetate toobtain 6. The filtrate was evaporated to afford 6 as a white solid.

The spectral data for compound 6 was:

¹H NMR (500 MHz, CDCl₃) δ 7.90 (d, 1H), 7.60 (d, 2H), 7.5-7.3 (m, 5H),6.30 (d, 2H), 4.5 (s, 2H), 2.3 (s, 2H).

A solution of 6 (2.79 mmol, 1.00 g) and p-nitrophenyl chloroformate(5.56 mmol, 1.12 g) was cooled to 0° C. Triethylamine (14.3 mmol, 2.0mL) was added and the solution was stirred for 15 minutes and pouredinto ammonium hydroxide. The solution mixture was poured into water andextracted with methylene chloride. The organic extract was washed withsaturated aqueous sodium bicarbonate, dried, and evaporated in vacuo toafford 7 (730 mg, 65%) as a white solid.

EXAMPLE 2 Synthesis of p38 Inhibitor Prodrugs 9 and 10

A mixture of 8 (1.0 g, 2.30 mmol) and N,N-dimethylformamide dimethylacetal (1.01 g, 6.91 mmol) in 10 mL of toluene was heated to 80° C. for20 minutes. The resulting solution was cooled to room temperature.Normal workup followed by chromatography on silica gel (hexane/EtOAc:10/4) gave amidine 9 (compound 101 of Table 1) as a white solid. Thespectral data for compound 9 was: ¹H NMR (500 MHz, CDCl₃) δ 8.3 (s, 1H),7.7 (d, 1H), 7.5-7.4 (m, 1H), 7.1-7.0 (m, 1H), 6.95-6.85 (t, 2H),6.85-6.75 (m, 1H), 6.45-6.4 (d, 1H), 6.2 (s, 1H), 4.95 (s, 2H), 3.05 (s,3H), 2.95 (s, 3H).

A mixture of 8 (1.0 g, 2.30 mmol) and N,N-dimethylformamide dimethylacetal (3.3 g, 22.4 mmol) in 10 mL of toluene was heated to 80° C. for90 minutes. The resulting solution was cooled to room temperature.Normal workup followed by chromatography on silica gel (hexane/EtOAc:2/1) gave bis-amidine 10 (compound 107 of Table 1) as a white solid. Thespectral data for compound 10 was: ¹H NMR (500 MHz, CDCl₃) δ 8.4 (s,1H), 8.3 (s, 1H), 8.05-7.95 (s, 1H), 7.15-7.05 (m, 2H), 6.85-6.75 (t,2H), 6.75-6.65 (m, 4H), 4.95 (s, 2H), 3.0-2.95 (d, 9H), 2.65 (s, 3H).

EXAMPLE 3 Synthesis of p38 Inhibitor Prodrug 13

To a mixture of 6 (1.25 gm, 3.35 mmol) and 4-nitrophenyl chloroformate(0.81 gm, 4.02 mmol) in tetrahydrofuran (30 mL) was added triethylamine(1.16 mL, 8.38 mmol) dropwise at 0° C. The resulting slurry was allowedto stir at 0° C. for 30 minutes. Ethanolamine (0.6 mL, 10.0 mmol) wasadded and the solution was stirred at 0° C. for 30 minutes. Normalwork-up followed by chromatography on silica (hexane/acetone: 10/4) gave11 (1.03 gm, 2.23 mmol) as a white solid. ¹H NMR (500 MHz, CDCl₃) 7.75(d, 1H), 7.65-7.55 (m, 2H), 7.5-7.4 (m, 1H), 7.25-7.15 (t, 2H),7.15-7.05 (t, 2H), 6.4 (d, 1H), 5.2-5.1 (bs, 1H), 5.15 (s, 2H),3.75-3.65 (t, 2H), 3.4-3.3 (m, 2H).

A mixture of 11 (1.03 gm, 2.23 mmol), (L)-BOC-Val-OH (0.97 gm, 4.46mmol), and 1-(3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloridein methylene chloride (30 mL) was stirred at room temperature for 1.5hours. Normal work-up followed by chromatography on silica(hexane/acetone: 10/4) gave Val deriv. 12 (1.38 gms, 2.09 mmol) as awhite solid. ¹H NMR (500 MHz, CDCl₃) 7.75 (d, 1H), 7.65-7.55 (m, 2H),7.5-7.4 (m, 1H), 7.25-7.15 (t, 2H), 7.15-7.05 (t, 2H), 6.4 (d, 1H),5.40-5.35 (bs, 1H), 5.05 (s, 2H), 5.00-4.95 (d, 1H), 4.4-4.3 (m, 1H),4.25-4.15 (m, 1H), 4.15-4.05 (m, 1H), 3.55-3.45 (m, 2H), 2.15-2.05 (m,1H), 1.45 (s, 9H), 1.0-0.85 (m, 6H).

To a solution of 12 (1.38 gms, 2.09 mmol) in methylene chloride (20 mLs)was added trifluoroacetic acid (10 mLs). The solution was allowed tostir at room temperature for 1 hour. Normal work-up gave a white solidthat was converted to its hydrochloride salt to give 13 (compound 111 ofTable 2; 0.61 gms, 1.02 mmol) as a white solid. The spectral data forcompound 13 was: ¹H NMR (500 MHz, CDCl₃) 7.65 (d, 1H), 7.55-7.45 (m,2H), 7.4-7.3 (m, 1H), 7.15-7.05 (m, 2H), 7.05-6.95 (m, 2H), 6.35 (d,1H), 5.05-5.00 (bs, 1H), 4.95 (s, 2H), 4.15-4.05 (m, 2H), 3.45-3.25 (m,2H), 3.2 (s, 1H), 1.95-1.85 (m, 1H), 0.90-0.75 (m, 6H).

EXAMPLE 4 Cloning of p38 Kinase in Insect Cells

Two splice variants of human p38 kinase, CSBP1 and CSBP2, have beenidentified. Specific oligonucleotide primers were used to amplify thecoding region of CSBP2 cDNA using a HeLa cell library (Stratagene) as atemplate. The polymerase chain reaction product was cloned into thepET-15b vector (Novagen). The baculovirus transfer vector,pVL-(His)6-p38 was constructed by subcloning a XbaI-BamHI fragment ofpET15b-(His)6-p38 into the complementary sites in plasmid pVL1392(Pharmingen).

The plasmid pVL-(His)6-p38 directed the synthesis of a recombinantprotein consisting of a 23-residue peptide (MGSSHHHHHHSSGLVPRGSHMLE,where LVPRGS represents a thrombin cleavage site) fused in frame to theN-terminus of p38, as confirmed by DNA sequencing and by N-terminalsequencing of the expressed protein. Monolayer culture of Spodopterafrugiperda (Sf9) insect cells (ATCC) was maintained in TNM-FH medium(Gibco BRL) supplemented with 10% fetal bovine serum in a T-flask at 27°C. Sf9 cells in log phase were co-transfected with linear viral DNA ofAutographa califonica nuclear polyhedrosis virus (Pharmingen) andtransfer vector pVL-(His)6-p38 using Lipofectin (Invitrogen). Theindividual recombinant baculovirus clones were purified by plaque assayusing 1% low melting agarose.

EXAMPLE 5 Expression and Purification of Recombinant p38 Kinase

Trichoplusia ni (Tn-368) High-Five™ cells (Invitrogen) were grown insuspension in Excel-405 protein free medium (JRH Bioscience) in a shakerflask at 27° C. Cells at a density of 1.5×10⁶ cells/ml were infectedwith the recombinant baculovirus described above at a multiplicity ofinfection of 5. The expression level of recombinant p38 was monitored byimmunoblotting using a rabbit anti-p38 antibody (Santa CruzBiotechnology). The cell mass was harvested 72 hours after infectionwhen the expression level of p38 reached its maximum.

Frozen cell paste from cells expressing the (His)₆-tagged p38 was thawedin 5 volumes of Buffer A (50 mM NaH₂PO₄ pH 8.0, 200 mM NaCl, 2 mMβ-Mercaptoethanol, 10% Glycerol and 0.2 mM PMSF). After mechanicaldisruption of the cells in a microfluidizer, the lysate was centrifugedat 30,000×g for 30 minutes. The supernatant was incubated batchwise for3-5 hours at 4° C. with Talon™ (Clontech) metal affinity resin at aratio of 1 ml of resin per 2-4 mgs of expected p38. The resin wassettled by centrifugation at 500×g for 5 minutes and gently washedbatchwise with Buffer A. The resin was slurried and poured into a column(approx. 2.6×5.0 cm) and washed with Buffer A+5 mM imidazole.

The (His)₆-p38 was eluted with Buffer A+100 mM imidazole andsubsequently dialyzed overnight at 4° C. against 2 liters of Buffer B,(50 mM HEPES, pH 7.5, 25 mM β-glycerophosphate, 5% glycerol, 2 mM DTT).The His₆ tag was removed by addition of at 1.5 units thrombin(Calbiochem) per mg of p38 and incubation at 20° C. for 2-3 hours. Thethrombin was quenched by addition of 0.2 mM PMSF and then the entiresample was loaded onto a 2 ml benzamidine agarose (AmericanInternational Chemical) column.

The flow through fraction was directly loaded onto a 2.6×5.0 cmQ-Sepharose (Pharmacia) column previously equilibrated in Buffer B+0.2mM PMSF. The p38 was eluted with a 20 column volume linear gradient to0.6M NaCl in Buffer B. The eluted protein peak was pooled and dialyzedovernight at 4° C. vs. Buffer C (50 mM HEPES pH 7.5, 5% glycerol, 50 mMNaCl, 2 mM DTT, 0.2 mM PMSF).

The dialyzed protein was concentrated in a Centriprep (Amicon) to 3-4 mland applied to a 2.6×100 cm Sephacryl S-100HR (Pharmacia) column. Theprotein was eluted at a flow rate of 35 ml/hr. The main peak was pooled,adjusted to 20 mM DTT, concentrated to 10-80 mgs/ml and frozen inaliquots at −70° C. or used immediately.

EXAMPLE 6 Activation of p38

p38 was activated by combining 0.5 mg/ml p38 with 0.005 mg/ml DD-doublemutant MKK6 in Buffer B+10 mM MgCl₂, 2 mM ATP, 0.2 mM Na₂VO₄ for 30minutes at 20° C. The activation mixture was then loaded onto a 1.0×10cm MonoQ column (Pharmacia) and eluted with a linear 20 column volumegradient to 1.0 M NaCl in Buffer B. The activated p38 eluted after theADP and ATP. The activated p38 peak was pooled and dialyzed againstbuffer B+0.2 mM Na₂VO₄ to remove the NaCl. The dialyzed protein wasadjusted to 1.1M potassium phosphate by addition of a 4.0M stocksolution and loaded onto a 1.0×10 cm HIC (Rainin Hydropore) columnpreviously equilibrated in Buffer D (10% glycerol, 20 mMβ-glycerophosphate, 2.0 mM DTT)+1.1MK₂HPO₄. The protein was eluted witha 20 column volume linear gradient to Buffer D+50 mM K₂HPO₄. The doublephosphorylated p38 eluted as the main peak and was pooled for dialysisagainst Buffer B+0.2 mM Na₂VO₄. The activated p38 was stored at −70° C.

EXAMPLE 7 p38 Inhibition Assays

A. Inhibition of Phosphorylation of EGF Receptor Peptide

This assay was carried out in the presence of 10 mM MgCl₂, 25 mMβ-glycerophosphate, 10% glycerol and 100 mM HEPES buffer at pH 7.6. Fora typical IC₅₀ determination, a stock solution was prepared containingall of the above components and activated p38 (5 nM). The stock solutionwas aliquotted into vials. A fixed volume of DMSO or inhibitor in DMSO(final concentration of DMSO in reaction was 5%) was introduced to eachvial, mixed and incubated for 15 minutes at room temperature. EGFreceptor peptide, KRELVEPLTPSGEAPNQALLR, a phosphoryl acceptor inp38-catalyzed kinase reaction (1), was added to each vial to a finalconcentration of 200 μM. The kinase reaction was initiated with ATP (100μM) and the vials were incubated at 30° C. After 30 minutes, thereactions were quenched with equal volume of 10% trifluoroacetic acid(TFA).

The phosphorylated peptide was quantified by HPLC analysis. Separationof phosphorylated peptide from the unphosphorylated peptide was achievedon a reverse phase column (Deltapak, 5 μm, C18 100D, Part no. 011795)with a binary gradient of water and acteonitrile, each containing 0.1%TFA. IC₅₀ (concentration of inhibitor yielding 50% inhibition) wasdetermined by plotting the percent (%) activity remaining againstinhibitor concentration.

B. Inhibition of ATPase Activity

This assay is carried out in the presence of 10 mM MgCl₂, 25 mMβ-glycerophosphate, 10% glycerol and 100 mM HEPES buffer at pH 7.6. Fora typical Ki determination, the Km for ATP in the ATPase activity ofactivated p38 reaction is determined in the absence of inhibitor and inthe presence of two concentrations of inhibitor. A stock solution isprepared containing all of the above components and activated p38 (60nM). The stock solution is aliquotted into vials. A fixed volume of DMSOor inhibitor in DMSO (final concentration of DMSO in reaction was 2.5%)is introduced to each vial, mixed and incubated for 15 minutes at roomtemperature. The reaction is initiated by adding various concentrationsof ATP and then incubated at 30° C. After 30 minutes, the reactions arequenched with 50 μl of EDTA (0.1 M, final concentration), pH 8.0. Theproduct of p38 ATPase activity, ADP, is quantified by HPLC analysis.

Separation of ADP from ATP is achieved on a reversed phase column(Supelcosil, LC-18, 3 μm, part no. 5-8985) using a binary solventgradient of following composition: Solvent A-0.1 M phosphate buffercontaining 8 mM tetrabutylammonium hydrogen sulfate (Sigma Chemical Co.,catalogue no. T-7158), Solvent B-Solvent A with 30% methanol.

Ki is determined from the rate data as a function of inhibitor and ATPconcentrations.

p38 inhibitors of this invention will inhibit the ATPase activity ofp38.

C. Inhibition of IL-1, TNF, IL-6 and IL-8 Production in LPS-StimulatedPBMCs

Inhibitors were serially diluted in DMSO from a 20 mM stock. At least 6serial dilutions were prepared. Then 4× inhibitor stocks were preparedby adding 4 μl of an inhibitor dilution to 1 ml of RPMI1640 medium/10%fetal bovine serum. The 4× inhibitor stocks contained inhibitor atconcentrations of 80 μM, 32 μM, 12.8 μM, 5.12 μM, 2.048 μM, 0.819 μM,0.328 μM, 0.131 μM, 0.052 μM, 0.021 μM etc. The 4× inhibitor stocks werepre-warmed at 37° C. until use.

Fresh human blood buffy cells were separated from other cells in aVacutainer CPT from Becton & Dickinson (containing 4 ml blood and enoughDPBS without Mg²⁺/Ca²⁺ to fill the tube) by centrifugation at 1500×g for15 min. Peripheral blood mononuclear cells (PBMCs), located on top ofthe gradient in the Vacutainer, were removed and washed twice withRPMI1640 medium/10% fetal bovine serum. PBMCs were collected bycentrifugation at 500×g for 10 min. The total cell number was determinedusing a Neubauer Cell Chamber and the cells were adjusted to aconcentration of 4.8×10⁶ cells/ml in cell culture medium (RPMI1640supplemented with 10% fetal bovine serum).

Alternatively, whole blood containing an anti-coagulant was useddirectly in the assay.

100 μl of cell suspension or whole blood were placed in each well of a96-well cell culture plate. Then 50 μl of the 4× inhibitor stock wasadded to the cells. Finally, 50 μl of a lipopolysaccharide (LPS) workingstock solution (16 ng/ml in cell culture medium) was added to give afinal concentration of 4 ng/ml LPS in the assay. The total assay volumeof the vehicle control was also adjusted to 200 μl by adding 50 μl cellculture medium. The PBMC cells or whole blood were then incubatedovernight (for 12-15 hours) at 37° C./5% CO₂ in a humidified atmosphere.

The next day the cells were mixed on a shaker for 3-5 minutes beforecentrifugation at 500×g for 5 minutes. Cell culture supernatants wereharvested and analyzed by ELISA for levels of IL-1β (R & D Systems,Quantikine kits, #DBL50), TNF-α (BioSource, #KHC3012), IL-6 (Endogen,#EH2-IL6) and IL-8 (Endogen, #EH2-IL8) according to the instructions ofthe manufacturer. The ELISA data were used to generate dose-responsecurves from which IC50 values were derived.

Results for the kinase assay (“kinase”; subsection A, above), IL-1, andTNF in LPS-stimulated PBMC's (“cell”) and IL-1, TNF, and IL-6 in wholeblood (“WB”) for various p38 inhibitors of this invention are shown inTable 7 below: TABLE 7 Kinase Cell IL-1 Cell TNF WB IL-1 WB TNF WB IL-6Compound M.W. IC50 (uM) IC50 (uM) IC50 (uM) IC50 (uM) IC50 (uM) IC50(uM) 13 559.55 0.031 0.012 0.022 0.140 0.055 0.083 9 489.43 1.0 0.050.05 12.2 20.0 11.0 10 544.51 5.0 2.2 4.3 0.8

Other p38 inhibitors of this invention will also inhibit phosphorylationof EGF receptor peptide, and will inhibit the production of IL-1, TNFand IL-6, as well as IL-8, in LPS-stimulated PBMCs or in whole blood.

D. Inhibition of IL-6 and IL-8 Production in IL-1-Stimulated PBMCs

This assay is carried out on PBMCs exactly the same as above except that50 μl of an IL-1b working stock solution (2 ng/ml in cell culturemedium) is added to the assay instead of the (LPS) working stocksolution.

Cell culture supernatants are harvested as described above and analyzedby ELISA for levels of IL-6 (Endogen, #EH2-IL6) and IL-8 (Endogen,#EH2-IL8) according to the instructions of the manufacturer. The ELISAdata are used to generate dose-response curves from which IC50 valueswere derived.

E. Inhibition of LPS-Induced Prostaglandin Endoperoxide Synthase-2(PGHS-2, or COX-2) Induction in PBMCs

Human peripheral mononuclear cells (PBMCs) are isolated from fresh humanblood buffy coats by centrifugation in a Vacutainer CPT (Becton &Dickinson). 15×10⁶ cells are seeded in a 6-well tissue culture dishcontaining RPMI 1640 supplemented with 10% fetal bovine serum, 50 U/mlpenicillin, 50 μg/ml streptomycin, and 2 mM L-glutamine. Compounds areadded at 0.2, 2.0 and 20 μM final concentrations in DMSO. LPS is thenadded at a final concentration of 4 ng/ml to induce enzyme expression.The final culture volume is 10 ml/well.

After overnight incubation at 37° C., 5% CO₂, the cells are harvested byscraping and subsequent centrifugation, the supernatant is removed, andthe cells are washed twice in ice-cold DPBS (Dulbecco's phosphatebuffered saline, BioWhittaker). The cells are lysed on ice for 10 min in50 μl cold lysis buffer (20 mM Tris-HCl, pH 7.2, 150 mM NaCl, 1%Triton-X-100, 1% deoxycholic acid, 0.1% SDS, 1 mM EDTA, 2% aprotinin(Sigma), 10 μg/ml pepstatin, 10 μg/ml leupeptin, 2 mM PMSF, 1 mMbenzamidine, 1 mM DTT) containing 1 μl Benzonase (DNAse from Merck). Theprotein concentration of each sample is determined using the BCA assay(Pierce) and bovine serum albumin as a standard. Then the proteinconcentration of each sample is adjusted to 1 mg/ml with cold lysisbuffer. To 100 μl lysate an equal volume of 2×SDS PAGE loading buffer isadded and the sample is boiled for 5 min. Proteins (30 μg/lane) aresize-fractionated on 4-20% SDS PAGE gradient gels (Novex) andsubsequently transferred onto nitrocellulose membrane by electrophoreticmeans for 2 hours at 100 mA in Towbin transfer buffer (25 mM Tris, 192mM glycine) containing 20% methanol. After transfer, the membrane ispretreated for 1 hour at room temperature with blocking buffer (5%non-fat dry milk in DPBS supplemented with 0.1% Tween-20) and washed 3times in DPBS/0.1% Tween-20. The membrane is incubated overnight at 4°C. with a 1:250 dilution of monoclonal anti-COX-2 antibody (TransductionLaboratories) in blocking buffer. After 3 washes in DPBS/0.1% Tween-20,the membrane is incubated with a 1:1000 dilution of horseradishperoxidase-conjugated sheep antiserum to mouse Ig (Amersham) in blockingbuffer for 1 h at room temperature. Then the membrane is washed again 3times in DPBS/0.1% Tween-20. An ECL detection system (SuperSignal™CL-HRP Substrate System, Pierce) is used to determine the levels ofexpression of COX-2.

EXAMPLE 8 ZAP70 Inhibition Assay

The activity of ZAP 70 is measured by determining the phosphorylationpoly E4Y (Sigma Chemicals, St Louis Mo.) with γ-³³P ATP (NEN, Boston,Mass.). Reactions are carried out at room temperature in a buffercontaining 100 mM HEPES, pH 7.5, 10 mM MgCl₂, 25 mM NaCl, 1 mM DTT and0.01% BSA. Final concentrations of ZAP70 and poly E4Y are 20 nM and 5 μMrespectively. Test compounds in DMSO (final concentration of compoundswas 30 μM in 1.5% DMSO) are added to the reaction mixture containing theabove-described components. The reaction is initiated by addition ofγ-³³P ATP (final concentration 20 μM, specific activity=0.018 Ci/mmol).The reaction is allowed to proceed for 12 minutes and then is quenchedby the addition of 10% TCA containing 200 mM ATP. The quenched reactionis harvested onto GF/C glass fiber filter plates (Packard, Meriden,Conn.) using a Tomtec 9600 cell harvester (Tomtec, Hamden, Conn.). Theplates are washed with 5% TCA containing 1 mM ATP and water. 50 μl ofscintillation fluid is added to the plates, which are then counted usinga Packard scintillation counter (Packard, Meriden, Conn.). IC50 valuesfor inhibitory compounds were determined using the same assay at aseries of compound concentrations.

While we have hereinbefore presented a number of embodiments of thisinvention, it is apparent that our basic construction can be altered toprovide other embodiments which utilize the methods of this invention.

1. A compound of the formula:

wherein each of Q₁ and Q₂ are independently selected from a phenyl or5-6 membered aromatic heterocyclic ring system, or a 8-10 memberedbicyclic ring system comprising aromatic carbocyclic rings, aromaticheterocyclic rings or a combination of an aromatic carbocyclic ring andan aromatic heterocyclic ring; wherein the rings that make up Q₁ aresubstituted with 1 to 4 substituents, each of which is independentlyselected from halo; C₁-C₃ alkyl optionally substituted with NR′₂, OR′,CO₂R′ or CONR′₂; O—(C₁-C₃)-alkyl optionally substituted with NR′₂, OR′,CO₂R′ or CONR′₂; NR′₂; OCF₃; CF₃; NO₂; CO₂R′; CONR′; SR′; S(O₂)N(R′)₂;SCF₃; CN; N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴;N(R′)R⁴; N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; or N═C—N(R′)₂; wherein therings that make up Q₂ are optionally substituted with up to 4substituents, each of which is independently selected from halogen;C₁-C₃ straight or branched alkyl optionally substituted with R′, NR′₂,OR′, CO₂R′, S(O₂)N(R′)₂, N═C—N(R′)₂, R³, O—P(O₃)H₂, or CONR′₂;O—(C₁-C₃)-alkyl; O—(C₁-C₃)-alkyl optionally substituted with NR′₂, OR′,CO₂R′, S(O₂)N(R′)₂, N═CR′—N(R′)₂, R³, OP(O₃)H₂, or CONR′₂; NR′₂; OCF₃;CF₃; NO₂; CO₂R′; CONR′₂; R³; OR³; NR³ ₂; SR³; C(O)R³; C(O)N(R′)R³;C(O)OR³; SR′; S(O₂)N(R′)₂; SCF₃; N═CR′—N(R′)₂; OR⁴; O—CO₂R⁴;N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴;N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; K; or CN; wherein each R′ isindependently selected from hydrogen; (C₁-C₃)-alkyl; (C₂-C₃)-alkenyl oralkynyl; phenyl or phenyl substituted with 1 to 3 substituentsindependently selected from halo, methoxy, cyano, nitro, amino, hydroxy,methyl or ethyl; or a 5-6 membered heterocyclic ring system optionallysubstituted with 1 to 3 substituents independently selected from halo,methoxy, cyano, nitro, amino, hydroxy, methyl or ethyl; wherein each Ris independently selected from hydrogen, —R², —N(R²)₂, —OR², SR²,—C(O)—N(R²)₂, —S(O₂)—N(R²)₂, —C(O)—OR² or —C(O)R² wherein two adjacent Rare optionally bound to one another and, together with each Y to whichthey are respectively bound, form a 4-8 membered carbocyclic orheterocyclic ring; wherein each R² is independently selected fromhydrogen; or (C₁-C₃)-alkyl or (C₁-C₃)-alkenyl, each optionallysubstituted with —N(R′)₂, —OR′, SR′, —O—C(O)—N(R′)₂, —C(O)—N(R′)₂,—S(O₂)—N(R′)₂, —C(O)—OR′, —NSO₂R⁴, —NSO₂R³, —C(O)N(R′)(R³), —NC(O)R⁴,—N(R′)(R³), —N(R′)(R⁴), —C(O)R³, —C(O)N(R′)(R⁴), —N(R⁴)₂, —C(O)N═C(NH)₂or R³; wherein each R³ is independently selected from 5-8 memberedaromatic or non-aromatic carbocyclic or heterocyclic ring systems eachoptionally substituted with R′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or —K; oran 8-10 membered bicyclic ring system comprising aromatic carbocyclicrings, aromatic heterocyclic rings or a combination of an aromaticcarbocyclic ring and an aromatic heterocyclic ring each optionallysubstituted with R′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or —K; wherein eachR⁴ is independently selected from R′; (C₁-C₇)-straight or branched alkyloptionally substituted with R′, N(R′)₂, OR′, CO₂R′, CON(R′)₂, SO₂N(R′)₂or SO₂N(R⁵)₂; or a 5-6 membered carbocyclic or heterocyclic ring systemoptionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, SO₂N(R′)₂ orSO₂N(R⁵)₂; wherein each R⁵ is independently selected from hydrogen, (C₁-C₃)-alkyl, or (C₁-C₃)-alkenyl; each optionally substituted with—N(R′)₂, —OR′, SR′, —C(O)—N(R′)₂, —S(O₂)—N(R′)₂, —C(O)—OR′,—N—S(O₂)(R′), —NSO₂R⁶, —C(O)N(R′)(R⁶), —NC(O)R′, —N(R′)(R⁶), —C(O)R⁶,—C(O)N═C(NH)₂ or R⁶; wherein each R⁶ is independently selected from 5-8membered aromatic or non-aromatic carbocyclic or heterocyclic ringsystems each optionally substituted with R′, —C(O)R′ or —C(O)OR′; or an8-10 membered bicyclic ring system comprising aromatic carbocyclicrings, aromatic heterocyclic rings or a combination of an aromaticcarbocyclic ring and an aromatic heterocyclic ring each optionallysubstituted with R′, —C(O)R′ or C(O)OR′; wherein R⁷ is selected from H,halogen, or a (C₁-C₃) straight chain or branched alkyl; wherein ach Y isindependently selected from N or C. If either Y is N, then R or Uattached to Y is a lone pair of electrons; wherein Z is CH, N, C(OCH₃),C(CH₃), C(NH₂), C(OH) or C(F); wherein each U is independently selectedfrom R or J; wherein each J is independently selected from a (C₁-C₄)straight chain or branched alkyl derivative substituted with T; whereineach T is independently selected from either O(V) or N(H)(V); whereineach V is independently selected from C(O)N═C(R)(N(R)₂), wherein the twogeminal R on the nitrogen are optionally bound to one another to form a4-8 membered carbocyclic or heterocyclic ring; wherein each K isindependently selected from a (C₁-C₄) straight chain or branched alkylderivative substituted with D, or —OP(O)(OH)₂; wherein each D isindependently selected from either enantiomer of

wherein each M is independently selected from either O or NH; whereineach G is independently selected from NH₂, OH, or H; wherein each R₈ isindependently selected from H, OH, C(O)OH, (C₁-C₇)-straight or branchedalkyl optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, orSO₂N(R⁵)₂; or a 5-6 membered carbocyclic, heterocyclic or heteroarylring system optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, orSO₂N(R⁵)₂; wherein G and R₈ are optionally bound to one another to forma ring.
 2. The compound according to claim 1, wherein Q₁ is selectedfrom phenyl or pyridyl containing 1 to 3 substituents independentlyselected from chloro, fluoro, bromo, —CH₃, —OCH₃, —OH, —CF₃, —OCF₃,—O(CH₂)₂CH₃, NH₂, 3,4-methylenedioxy, —N(CH₃)₂, —NH—S(O)₂-phenyl,—NH—C(O)O—CH₂-4-pyridine, —NH—C(O)CH₂-morpholine, —NH—C(O)CH₂—N(CH₃)₂,—NH—C(O)CH₂-piperazine, —NH—C(O)CH₂-pyrrolidine,—NH—C(O)C(O)-morpholine, —NH—C(O)C(O)-piperazine,—NH—C(O)C(O)-pyrrolidine, —O—C(O)CH₂—N(CH₃)₂, or —O—(CH₂)₂—N(CH₃)₂ andwherein at least one of said substituents is in the ortho position. 3.The compound according to claim 2, wherein Q₁ contains at least twosubstituents, both of which are in the ortho position.
 4. The compoundaccording to claim 2, wherein Q₁ is selected from:


5. The compound according to claim 4, wherein Q₁ is selected from2-fluoro-6-trifluoromethylphenyl, 2,6-difluorophenyl,2,6-dichlorophenyl, 2-chloro-4-hydroxyphenyl, 2-chloro-4-aminophenyl,2,6-dichloro-4-aminophenyl, 2,6-dichloro-3-aminophenyl,2,6-dimethyl-4-hydroxyphenyl, 2-methoxy-3,5-dichloro-4-pyridyl,2-chloro-4,5methylenedioxy phenyl, or2-chloro-4-(N-2-morpholino-acetamido)phenyl.
 6. The compound accordingto claim 1, wherein Q₂ is selected from phenyl, pyridyl or naphthyl andwherein Q₂ optionally contains up to 3 substituents, each of which isindependently selected from chloro, fluoro, bromo, methyl, ethyl,isopropyl, —OCH₃, —OH, —NH₂, —CF₃, —OCF₃, —SCH₃, —OCH₃, —C(O)OH,—C(O)OCH₃, —CH₂NH₂, —N(CH₃)₂, —CH₂-pyrrolidine and —CH₂OH.
 7. Thecompound according to claim 6, wherein Q₂ is selected from:

unsubstituted 2-pyridyl or unsubstituted phenyl.
 8. The compoundaccording to claim 7, wherein Q₂ is selected from phenyl,2-isopropylphenyl, 3,4-dimethylphenyl, 2-ethylphenyl, 3-fluorophenyl,2-methylphenyl, 3-chloro-4-fluorophenyl, 3-chlorophenyl,2-carbomethoxylphenyl, 2-carboxyphenyl, 2-methyl-4-chlorophenyl,2-bromophenyl, 2-pyridyl, 2-methylenehydroxyphenyl, 4-fluorophenyl,2-methyl-4-fluorophenyl, 2-chloro-4-fluorphenyl, 2,4-difluorophenyl,2-hydroxy-4-fluorphenyl or 2-methylenehydroxy-4-fluorophenyl,1-naphthyl, 3-chloro-2-methylenehydroxy, 3-chloro-2-methyl, or4-fluoro-2-methyl.
 9. The compound according to claim 1, wherein each Yis C.
 10. The compound according to claim 9, wherein the R attached to Yis independently selected from hydrogen or methyl.
 11. The compoundaccording to claim 1, wherein J is a 0-8 atom chain terminating in analcohol, amine, carboxylic acid, ester, amide, amidine or heterocycle.12. The compound according to claim 11, wherein J is selected from:


13. The compound according to claim 1 wherein K is selected from:


14. The compound according to claim 1, wherein the compound is selectedfrom any one of the compounds depicted in Tables 1-3.
 15. The compoundaccording to claim 1, wherein the compound is

wherein Ar is


16. The compound according to claim 1, wherein the compound is

wherein Ar is


17. The compound according to claim 1, wherein the compound is

wherein Ar is


18. The compound according to claim 1, wherein the compound is

wherein


19. The compound according to claim 1, wherein the compound is

wherein


20. A pharmaceutical composition comprising an amount of a compoundaccording to claim 1 effective to inhibit p38, and a pharmaceuticallyacceptable carrier. 21-33. (canceled)
 34. A pharmaceutical compositioncomprising an amount of a compound according to claim 1 effective toinhibit ZAP70, and a pharmaceutically acceptable carrier. 35-38.(canceled)